title: Characterization of the mechanism of inhibin α-subunit gene in mouse anterior pituitary cells by RNA interference.
abstract: Inhibin, a member of the transforming growth factor-β [TGF-β] superfamily, is a suppressor of follicle-stimulating hormone [FSH] release through pituitary-gonadal negative feedback loop to regulate follicular development. In this study, Inhibin α-subunit [Inha] gene was knocked down successfully in mice primary anterior pituitary cells at both transcriptional and translational levels by RNAi-Ready pSIREN-RetroQ-ZsGreen Vector mediated recombinant pshRNA vectors. The results indicated that inhibin silencing significantly promoted apoptosis by up-regulating Caspase-3, Bax and Bcl-2 genes without affecting p53 both at transcriptional and translational levels. Furthermore, it markedly impaired the progression of G1 phase of cell cycle and decreased the amount of cells in S phase [as detected by flow cytometry]. Inhibin silencing resulted in significant up-regulation of mRNA and protein expressions of Gondotropin releasing hormone receptors [GnRHR] and down-regulated mRNA levels of β-glycans with parellel change in the amount of its protein expression. Silencing of inhibin-a significantly increased [P<0.05] activin-β concentration without affecting FSH and LH levels in anterior pituitary cells. These findings revealed that up regulation of GnRH receptors by silencing inhibin a-subunit gene might increase the concentration of activin-β in the culture medium. Inhibin a silencing resulted in increased mRNA and protein expressions of inhibinβ which may demonstrate that both inhibin subunits co-participate in the regulation of reproductive events in anterior pituitary cells. This study concludes that inhibin is a broad regulatory marker in anterior pituitary cells by regulating apoptosis, cellular progression and simultaneously by vital fluctuations in the hormonal signaling.
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Laboratory of Animal Infectious Diseases, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, China ; Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education. College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, China.
authors.forename: Li
authors.initials: L
authors.lastname: Han
authors.forename: Canjie
authors.initials: C
authors.lastname: Wu
authors.forename: Hasan
authors.initials: H
authors.lastname: Riaz
authors.forename: Liya
authors.initials: L
authors.lastname: Bai
authors.forename: Jianguo
authors.initials: J
authors.lastname: Chen
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Zhen
authors.forename: Aizhen
authors.initials: A
authors.lastname: Guo
authors.forename: Liguo
authors.initials: L
authors.lastname: Yang
mesh_terms: D028341:Activins / Q000378:metabolism; D000818:Animals; D053148:Caspase 3 / Q000235:genetics; D049109:Cell Proliferation; D005260:Female; D005640:Follicle Stimulating Hormone / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics; D007265:Inhibins / Q000172:deficiency* / Q000235:genetics* / Q000378:metabolism; D007986:Luteinizing Hormone / Q000378:metabolism; D051379:Mice; D010903:Pituitary Gland, Anterior / Q000166:cytology* / Q000378:metabolism; D011509:Proteoglycans / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D011966:Receptors, LHRH / Q000235:genetics / Q000378:metabolism; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics; D012098:Reproduction; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D051028:bcl-2-Associated X Protein / Q000235:genetics
chemical_list: D011509:Proteoglycans; D019253:Proto-Oncogene Proteins c-bcl-2; D034741:RNA, Small Interfering; D011966:Receptors, LHRH; D018125:Receptors, Transforming Growth Factor beta; D016159:Tumor Suppressor Protein p53; C044147:activin B; D051028:bcl-2-Associated X Protein; C094377:inhibin-alpha subunit; D028341:Activins; C052617:Bcl2 protein, mouse; C071736:betaglycan; D007265:Inhibins; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D053148:Caspase 3